development of a sybr green real time multiplex rt-pcr technique for simultaneous detection of hcv and gbv-c co-infection in plasma samples

development of a sybr green real time multiplex rt-pcr technique for simultaneous detection of hcv and gbv-c co-infection in plasma samples

seyed hossein mousavi fard, shahin merat, kiana shahzamani, reza ghanbari, neda yahoo

Abstract

background:accumulative research is in progress to clarify clinical aspects of gbv-c. the possibility of interaction between hcv and gbv-c as well as its consequence on development of liver diseases is the most important clinical aspect which encourages researchers to develop a rapid and cost effective technique for simultaneous detection of both viruses.methods: in this study, a sybr green real time multiplex rt-pcr technique as a new economical and sensitive method was designed and validated for simultaneous detection of hcv/gbv-c in hcv positive plasma samples. sybr green real time rt-pcr technique optimization was performed separately for each virus. multiplex pcr was established next. standard sera with known concentrations of hcv rna and dual hcv/gbv-c positive control samples along with negative control samples were used to validate the assay.results and conclusions: fifty six non cirrhotic hcv positive plasma samples [29 of genotype 3a and 27 of genotype 1a] were collected from patients before receiving treatment. 20.6% of genotype 3a and 18.7% of genotype 1a showed hcv/gbv-c co-infection. as a result, 19.6% of 56 samples had hcv/gbv-c co-infection that was compatible with other results from all over the world. sybr green real time multiplex rt-pcr technique can be used to detect hcv/gbv-c co-infection in plasma samples. furthermore, with application of this method more time and cost could be saved in clinical-research settings.